A novel serine/threonine kinase associated with thrombin receptor signaling in human platelet

一种与人血小板中凝血酶受体信号传导相关的新型丝氨酸/苏氨酸激酶

• 批准号: 08680680
• 负责人: IDO Masaru
• 金额: $ 1.6万
• 依托单位: Mie University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08680680/
• 关键词: Thrombin receptor; Protein kinase; Platelet; Proteim Kinase; トロンビンレセプター

Novel protein kinases that may associate with thrombin receptor were sought by a procedure based on the ability of these enzymes to undergo renaturation and catalyze the phosphorylation of a protein substrate fixed in a gel. We employed a fusion protein comprising of glutathione S-transferase and the substrate protein that corresponds to residues 375-421 of thrombin receptor (GST-TRCD). We report that platelets contain a serine/threonine protein kinase with molecular masse of about 33 kDa, which specifically bound to GST-TRCD.Treatment of platelets with thrombin increased the thrombin receptor-associated kinase (TRAK) activity by 700%. TRAK was also activated by a thrombin receptor agonist peptide, but not by hirudin-treated or diisopropylphosphate-inactivated thrombin. Activation of thrombin receptor resulted in phosphorylation of TRCD by TRAK in a Ca^<2+>-dependent way. In summary, our results provide evidence of a novel ligand-activated serine/threonine kinase that associates with the cytoplasmic domain of the thrombin receptor.

通过一种基于这些酶的复性和催化凝胶中固定的蛋白质底物的磷酸化能力的程序，寻找可能与凝血酶受体相关的新的蛋白激酶。我们使用了谷胱甘肽S转移酶与凝血酶受体第375-421残基的底物蛋白(GST-TRCD)的融合蛋白。我们报道了血小板含有一个分子质量约为33 kDa的丝氨酸/苏氨酸蛋白激酶，它与GST-TRCD特异性结合。凝血酶处理使血小板凝血酶受体相关激酶(TRAK)活性增加了700%。TRAK也可以被凝血酶受体激动肽激活，但不能被水飞蓟素治疗或磷酸二异丙酯灭活的凝血酶激活。凝血酶受体的激活导致TRAK以钙依赖的方式使TRCD磷酸化。总之，我们的结果提供了一种新的配体激活的丝氨酸/苏氨酸激酶的证据，它与凝血酶受体的胞浆结构域相关联。

项目成果

Takeya, H., Mori, T., Gabazza, E.C., Kuroda, K., Deguchi, H., Matsuura, E., Ichikawa, K., Koike, T.and Suzuki, K.: "Anti-beta2-glycoprotein I (beta2GPI) monoclonal antibodies with lupus anticoagulant-like activity enhance the beta2GPI binding to phospholi

Takeya, H.、Mori, T.、Gabazza, E.C.、Kuroda, K.、Deguchi, H.、Matsuura, E.、Ichikawa, K.、Koike, T. 和 Suzuki, K.：“抗 β2-糖蛋白

Ido M et al.: "Ca_<2+>-dependent activation of the 33-kDa protein Kinase transmits thrombin receptor signals in human platelets." Thromb Haemostas. 76. 439-443 (1996)

Ido M等人：“33-kDa蛋白激酶的Ca_2依赖性激活在人血小板中传递凝血酶受体信号。”

Ido M et al.: "Blood Coagulation,Fibrinolysis,and Platebts" Springer E.W.Davie & E. Kakeshita, 228 (1996)

Ido M 等人：“血液凝固、纤维蛋白溶解和血小板” Springer E.W.Davie

Ido, M.: "Blood Coagulation,Fibrinolysis,and Platelets." Davie EW,Kakeshita K,eds.Splinger-Verlag Tokyo, 228 (1996)

Ido, M.：“凝血、纤维蛋白溶解和血小板。”

Ido M et al: "Ca^<2+>-dependent activation of the 33-kDa protein kinase transmits thrombin receptor signals in human platelets." Thromb Haemostas. 76. 439-443 (1996)

Ido M 等人：“33-kDa 蛋白激酶的 Ca^2 依赖性激活在人血小板中传递凝血酶受体信号。”